基本信息
| 启动子: | CMV IE |
| 复制子: | pUC ori，F1 ori |
| 终止子: | SV40 poly（A） signal |
| 质粒分类: | 哺乳细胞载体;蛋白过表达质粒 |
| 质粒大小: | 5472bp |
| 原核抗性: | 氨苄青霉素Amp |
| 筛选标记: | 新霉素Neo |
| 克隆菌株: | DH5α |
| 培养条件: | 37℃，有氧，LB |
| 表达宿主: | 哺乳细胞 |
| 培养条件: | 37℃，有氧，LB |
| 诱导方式: | 无需诱导，瞬时表达 |
| 5'测序引物: | CMV-F: 5-CGCAAATGGGCGGTAGGCGTG-3 |
| 3'测序引物: | EBV-R:5-GTGGTTTGTCCAAACTCATC-3 |
质粒简介
pCI-neo Mammalian Expression Vector(pCI-neo 哺乳动物表达载体) 携带有人类巨细胞病毒(CMV) 的即刻早期增强子/ 启动子区域，来促进哺乳动物细胞中插入克隆DNA 的组成型表达。此载体也带有新霉素磷酸转移酶基因, 一种哺乳动物细胞的选择性标志物。通过G-418 抗性筛选转染细胞，pCI-neo 载体能够用于瞬时或稳定表达。
质粒图谱
质粒序列
LOCUS Exported 5472 bp ds-DNA circular SYN 06-11-2015
DEFINITION .
ACCESSION .
VERSION .
KEYWORDS Untitled
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5472)
AUTHORS admin
TITLE Direct Submission
JOURNAL Exported 2015-11-6 from MLCC
FEATURES Location/Qualifiers
source 1..5472
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 138..517
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 518..729
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
intron 890..1022
/note="chimeric intron"
/note="chimera between introns from human beta-globin and
immunoglobulin heavy chain genes"
promoter 1067..1085
/note="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(1141..1158)
/note="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
polyA_signal 1176..1297
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 1483..1938
/direction=RIGHT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2055..2412
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2263..2398
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 2463..3257
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3321..3369
/note="synthetic polyadenylation signal"
promoter 3675..3779
/gene="bla"
/note="AmpR promoter"
CDS 3780..4640
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/note="AmpR"
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 4811..5399
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 tcaatattgg ccattagcca tattattcat tggttatata gcataaatca atattggcta
61 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
121 aatatgaccg ccatgttggc attgattatt gactagttat taatagtaat caattacggg
181 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
241 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
301 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
361 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
421 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
481 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
541 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
601 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactg
661 cgatcgcccg ccccgttgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
721 agcagagctc gtttagtgaa ccgtcagatc actagaagct ttattgcggt agtttatcac
781 agttaaattg ctaacgcagt cagtgcttct gacacaacag tctcgaactt aagctgcagt
841 gactctctta aggtagcctt gcagaagttg gtcgtgaggc actgggcagg taagtatcaa
901 ggttacaaga caggtttaag gagaccaata gaaactgggc ttgtcgagac agagaagact
961 cttgcgtttc tgataggcac ctattggtct tactgacatc cactttgcct ttctctccac
1021 aggtgtccac tcccagttca attacagctc ttaaggctag agtacttaat acgactcact
1081 ataggctagc ctcgagaatt cacgcgtggt acctctagag tcgacccggg cggccgcttc
1141 cctttagtga gggttaatgc ttcgagcaga catgataaga tacattgatg agtttggaca
1201 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
1261 tttatttgta accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt
1321 tatgtttcag gttcaggggg agatgtggga ggttttttaa agcaagtaaa acctctacaa
1381 atgtggtaaa atccgataag gatcgatccg ggctggcgta atagcgaaga ggcccgcacc
1441 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat ggacgcgccc tgtagcggcg
1501 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
1561 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
1621 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
1681 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
1741 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
1801 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
1861 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa
1921 tattaacgct tacaatttcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc
1981 acaccgcata cgcggatctg cgcagcacca tggcctgaaa taacctctga aagaggaact
2041 tggttaggta ccttctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
2101 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
2161 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
2221 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
2281 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
2341 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
2401 ggcttttgca aaaagcttga ttcttctgac acaacagtct cgaacttaag gctagagcca
2461 ccatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
2521 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
2581 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
2641 tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
2701 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
2761 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
2821 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
2881 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
2941 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg
3001 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
3061 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
3121 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
3181 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
3241 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
3301 cctgccatca cgatggccgc aataaaatat ctttattttc attacatctg tgtgttggtt
3361 ttttgtgtga atcgatagcg ataaggatcc gcgtatggtg cactctcagt acaatctgct
3421 ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac
3481 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
3541 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
3601 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
3661 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
3721 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
3781 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
3841 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
3901 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
3961 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
4021 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
4081 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
4141 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
4201 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
4261 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
4321 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
4381 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
4441 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
4501 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
4561 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
4621 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
4681 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
4741 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
4801 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
4861 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
4921 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
4981 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
5041 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
5101 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
5161 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
5221 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
5281 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
5341 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
5401 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgg
5461 ctcgacagat ct
温馨提示：不可用于临床ZL。
